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Biomarker discovery for Parkinson’s disease

Biomarker discovery for Parkinson’s disease

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Piu CHAN Department of Neurology Neurobiology and GeriatricsBeijing Institute of GeriatricsXuanwu Hospital of Capital Medical University

＜正＞Parkinson’s disease（PD） is a chronic debilitating disease characterized by the selective and progressive loss of nigrostriatal dopamine *** mechanisms underlying the pathophysiology of this disease are not fully *** dysfunction,oxidative damage,excitotoxicity,excessive iron accumulation and inflammation are likely to play a role in cell *** it is believed that toxic environmental factors,along with a possible genetic susceptibility,may lead to disease *** of the most imperative and challenging issues in PD field is to establish a standard for early *** one hand,by the time patients with PD become symptomatic and seek medical treatment,60%-80%of the neurons in the substantia nigra pars compacta have been irreversibly lost,meaning the present clinical diagnostic means are not sensitive *** the other,although parkinson’s disease affects patients later in life,abundant evidence suggests the existence of a "preclinical" stage that could start years before a subject can be diagnosed clinically,when an individual appears normal but is developing extensive pathological changes in he ***,early detection of PD is realizable,and in turn will improve disease management and enable potential neuroprotective therapies to be introduced at a stage when they would provide the greatest *** addition,biomarkers would also be useful in monitoring the effectiveness of treatments for *** undertake the initiative for discovery of biomarkers for PD,efforts have been made to establish national consortium and community longitudinal cohorts using standardized protocols for collection of clinical information and tissue *** addition,non-human primate model for PD has also been established to mimic the pathological processes of PD for novel biomarker *** talk will be focused on the establishment of the above-mentioned cohorts and models and preliminary results.

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Kinesin superfamily molecular motors,KIFs:From intracellular transport to brain development,wiring and learning and memory

Kinesin superfamily molecular motors,KIFs:From intracellular...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Nobutaka Hirokawa Department of Cell Biology and Anatomy Graduate School of MedicineUniversity of Tokyo

＜正＞The intracellular transport is fundamental for neuronal morphogenesis and *** elucidate the mechanism we have identified and characterized kinesin superfamily proteins,KIFs,by molecular cell biology, molecular genetics,biophysics and structural *** transport cargoes such as synaptic vesicle precursors （KIF1A/KIF1B beta）,mitochondria（KIF1A alpha/KIF5）,NMDA receptor（KIF17）,AMPA receptor（KIF5） and mRNA with a large protein complex（KIF5） in the axon and *** recognize their own cargoes mostly through scaffold or adaptor protein complexes and release cargoes by phosphorylation of KIFs or through GTP hydrolysis of Rab protein in ***,we elucidated significant unexpected functions of ***2A is fundamental for neuronal migration and brain wiring by depolymerizing microtubules in growth cones and suppressing unnecessary elongation of axon ***4 controls the activity-dependent survival of neurons by regulating PARP-1 activity during brain ***3 suppresses tumorigenesis by transporting beta catenin to the plasma membrane. KIF26A controls development of enteric nervous system as a key suppressor of GDNF/Ret signaling ***, KIFs play a number of significant roles not only on various neuronal functions by transporting important cargos,but also on fundamental physiological processes such as brain wiring,suppression of tumorigenesis,and control of activity dependent neuronal survival and higher brain functions.

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The morphogens Shh and Wnts cooperate in axon guidance

The morphogens Shh and Wnts cooperate in axon guidance

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Esther T.Stoeckli Institute of Molecular Life Sciences University of ZurichSwitzerland

Commissural axons of the spinal cord are a widely used model to study molecular mechanisms of axon *** crossing the floor plate,the ventral midline of the spinal cord,axons of dorsolateral commissural neurons turn into the longitudinal axis and extend in rostral direction.A variety of cues have been identified for guidance towards and across the *** morphogen Sonic hedgehog(Shh) was identified as a chemoattractant for commissural axons that acts in parallel to Netrin(Charron et al,Cell 2003).After reaching the midline,commissural axons change their responsiveness to Shh by switching ***-interacting protein(Hhip) mediates a repulsive activity of Shh guiding post-crossing commissural axons rostrally,in accordance with the anteroposterior gradient of Shh in the floor plate(Bourikas et al,***.2005).In the mouse,Wnt4 expressed in an antero-posterior gradient was shown to attract post-crossing commissural axons rostrally(Lyuksyutova et al, Science 2003).We found that Wnts also act as axon guidance cues in the chicken spinal ***,in contrast to the mouse,Wnt transcription did not differ along the anteroposterior axis of the chicken spinal cord(Domanitskaya et al,***.2010).Rather,Wnt activity was regulated by a graded distribution of Sfrps(Secreted frizzledrelated proteins) that in turn appears to be shaped by the graded expression of ***,Shh affects post-crossing commissural axon guidance both directly and indirectly by regulating Wnt activity.

关键词： axon guidance morphogens Shh Wnt neural circuit formation

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hGaiR2与Cript的相互作用

hGaiR2与Cript的相互作用

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者：路雅静宫夏霓李天翊徐舒杨予涛徐志卿首都医科大学神经生物学系北京神经科学研究所北京神经再生修复研究重点实验室

目的甘丙肽是一个含29个氨基酸的神经肽,其广泛分布于中枢和外周组织,参与多种生理过程,如:摄食,情绪,记忆,神经生长等。目前已经克隆出三种甘丙肽受体,分别是GalR1,GalR2,GalR3,它们都属于G蛋白偶联受体,在同一物种中,各种GalR之间呈... 详细信息

目的甘丙肽是一个含29个氨基酸的神经肽,其广泛分布于中枢和外周组织,参与多种生理过程,如:摄食,情绪,记忆,神经生长等。目前已经克隆出三种甘丙肽受体,分别是GalR1,GalR2,GalR3,它们都属于G蛋白偶联受体,在同一物种中,各种GalR之间呈现很低序列相似性。本研究主要关注的是GalR2的功能及转运。方法酵母双杂交实验寻找可以与GalR2相互作用的蛋白。钓出的蛋白再经酵母双转验证,再次证实两种蛋白可以发生相互作用。CO-IP再次验证两种蛋白的可以发生相互作用。结果经过以上我们找到了Cript这个蛋白,Cript是一个有101个氨基酸的小分子量蛋白,其可与微管蛋白结合,帮助PSD95蛋白与微管的锚定。而Cript与GalR2的结合发挥什么作用,需要继续去探寻。目前已经克隆出Cript,并且用CO-IP的方法验证出Cript确实可以与GalR2发生相互作用。

关键词： GalR2 Cript 酵母双杂交

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Retinal dopaminergic cells,control of eye growth and the prevention of myopia

Retinal dopaminergic cells,control of eye growth and the pre...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Ian Morgan Regan Ashby ARC Centre of Excellence in Vision Science Research School of BiologyAustralian National UniversityCanberraAustralia Division of Preventive Ophthalmology Zhongshan Ophthalmic CenterSun Yat-Sen UniversityGuangzhouChina Division of Health Sciences University of CanberraCanberraAustralia

＜正＞Dopaminergic amacrine cells,although few in number,form a mosaic covering the entire *** release of dopamine is dependent on light intensity as well as the spatio-temporal properties of the light stimulus. The dopaminergic cells are,in part,driven by ON-bipolar cells,and are also under the control of reciprocal inhibitory interactions with photoreceptors（using the modulator melatonin） and another population of amacrine cells which uses enkephalins,neurotensin and somatostatin are *** rate of dopamine release is thus maintained at a low level in the dark,but increases with light intensity and spatio-temporal contrast in the *** appears to be involved in a large range of functions,including regulation of light adaptation,and horizontal cell *** animal models of experimental myopia,dopamine release is reduced,and dopamine agonists then are able to inhibit the excessive axial elongation underlying the development of *** studies on children have shown that children who spend more time outside are less likely to become myopic,possibly due to the higher light intensities encountered outdoors as compared to indoors during daylight *** phenomenon has been replicated in animals exposed to myopigenic stimuli,where exposure to bright lights also inhibits the development of myopia. This inhibition can be prevented by intraocular injection of D2-dopamine ***-based interventions are currently underway in Guangzhou to determine if increasing the amount of time that children are exposed to bright light outdoors,which should stimulate the rate of release of dopamine,will prevent the rapid development of myopia that currently occurs in children in *** studies are also in progress on the retinal pathways which mediate these effects,which may open up possibilities for pharmacological intervention.

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High throughput screening of novel inhibitors for TRPC4/C5 channels

High throughput screening of novel inhibitors for TRPC4/C5 c...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Michael X.Zhu Department of Integrative Biology and Pharmacology The University of Texas Health Science Center at HoustonHouston TexasUSA

Canonical transient receptor potential 4 and 5(TRPC 4 and TRPC5) are non-selective cation channels widely expressed in many mammalian *** of these channels causes membrane depolarization and intracellular Ca ***4 has been implicated in neurotransmitter release from thalamic interneurons, endothelial-dependent regulation of vascular tone,microvascular permeability,and excitation-contraction coupling of intestinal smooth muscles while TRPC5 has been shown to be important for neurite extension and growth cone morphology of hippocampal neurons and behavior responses to fear *** channels are thought to be responsible for afterdepolarization of cerebral cortex and hippocampal *** whole-cell patch clamp recording and fluorescence membrane potential measurements,we show that channel activation of TRPC4 and C5 are dependent on the co-stimulation of G- and G-coupled *** effect of G proteins appears to be mainly mediated through the activated form of G rather than Gβγ.Taking the advantage of G-induced TRPC4 activation and the robust Ca influx signal,we have developed a cell-based assay for high throughput screening of TRPC channel *** screening was performed at the Johns Hopkins Ion Channel Center against the Molecular Libraries Small Molecule Repository(MLSMR),yielding>1000 lead compounds as potential agonists,antagonists and allosteric *** have performed detailed characterizations on two antagonists,showing excellent selectivity to TRPC4/C5 as compared to other TRP channels and voltage-gated K,Na,and Ca *** compounds will be valuable tools for dissecting the mechanisms of regulation and physiological functions of these novel channels.

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Dynamics of synapse remodeling and glia motility in vivo

Dynamics of synapse remodeling and glia motility in vivo

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Junichi Nabekura Go Kato Kei Eto Kwan Sun Kim National Institute for Physiological Sciences(NIPS)

Recent advance in imaging techniques,*** MRI and PET,allows us to understand the activation of various brain ***,due to the spatial limitation of these approaches,it is difficult to observe more fine structures,*** and synapses in *** photon excitation of fluorescent molecules enables us to observe the fine structures and neuronal activity in the various organs and ***,we introduce two evidences of dynamics of neuronal structures and glia in the brain of living *** time imaging with two photon microscopy revealed that resting microglia,the primary immune effector cells in the brain,dynamically extend and retract their processes as if actively surveying the *** directly monitors the local synaptic states. Resting microglial processes make brief(～5 min) and direct contacts with neuronal synapses at a frequency of about once per *** transient cerebral ischemia the microglia-synapse contact periods were markedly prolonged ( hour),frequently followed by the disappearance of presynaptic *** striking finding of the synapse disappearance driven by microglial contact could contribute to an increased turnover of synaptic connections and remodeling of neuronal circuits receiving ischemic *** observation of the same neuronal structures of the mouse cortex over several months reveals the dynamic change of dendritic spines of somatosensory cortex in case of neuropathic pain induced by peripheral ligation of sciatic *** turnover in the S1 area corresponding to the injured paw markedly increased during an early developing phase of neuropathic pain and was restored in a late phase of neuropathic *** stable spines survived less following injury than controls,and new generated spine after the injury tend to preferentially survive,which displayed long-term potentiation-like enlargement over weeks. These results suggest that peripheral nerve injury induces rapid a

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Channelepsy:channelopathy in epilepsies

Channelepsy:channelopathy in epilepsies

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Shinichi HIROSE Department of Pediatrics School of Medicine Research Institute for the Molecular Pathomechanisms of EpilepsyFukuoka University

"Channlepsy" is a compounder of channelopathy—i.e.,disease resulting from channel dysfunction—and *** thus indicates epilepsy resulting from channel *** notion that some,if not all epilepsies,are channelopathies,is a result of recent molecular research into *** the notion,in fact,some 20 epilepsies have been recognized as "channelepsies".Such channelepsies include autosomal dominant nocturnal frontal lobe epilepsy(ADNFLE) resulting from mutations of neuronal nicotinic acetylcholine receptor (nAChR),which is a ligand gated ion channel,benign familial neonatal seizures(BFNS) from mutations of potassium channels,genetic epilepsy with febrile seizures plus(GEFS+) and Dravet syndrome from mutations of sodium channels and *** channelopathies have led to a new strategy for investigating the pathomechanisms of *** ion channels can be reconstituted in oocyte or cultivated cells using relevant cDNA expression systems, ion channel dysfunctions due to genetic abnormalities can be readily evaluated in vitro,enabling us to understand the molecular and neuroscientific pathomechanisms underlying various *** addition,this mutational information makes possible genetically engineered animal models for epilepsy(GEAM),which harbor mutations identified in human *** GEAM can represent molecular pathomechanisms more precisely than conventional animal models and their use has revealed several of the pathomechanisms underlying *** first pathomechanism identified was the imbalance between the neuronal excitatory and inhibitory systems;for example, mutations of KCNQ2,the gene encoding the potassium channel K7.2,cause *** KCNQ2 mutations were found to lead to loss of function of K7.2 in the in vitro system.K7.2,along with its counterpart K7.3,generates potassium current,called M-current,which plays an important role in the neuronal inhibitory ***,K7.2 or K7.3 dysfunction undermine

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皮层下缺血性血管性痴呆疾病机制的初探及肌肽对其作用的研究

皮层下缺血性血管性痴呆疾病机制的初探及肌肽对其作用的研究

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者：马婧张晨晖闫海静王熔融胡薇薇陈忠浙江大学药学院浙江大学医学院药理系浙江大学药学院浙江大学医学院药理系

目的血管性痴呆是目前继老年性痴呆之后排名第二的痴呆疾病。皮层下缺血性血管性痴呆是血管性痴呆最常见的亚型,但发病机制尚不清楚,并且至今尚无非常有效的治疗手段。肌肽是一种在动物体脑内广泛分布的天然二肽化合物,具有多种生物学... 详细信息

目的血管性痴呆是目前继老年性痴呆之后排名第二的痴呆疾病。皮层下缺血性血管性痴呆是血管性痴呆最常见的亚型,但发病机制尚不清楚,并且至今尚无非常有效的治疗手段。肌肽是一种在动物体脑内广泛分布的天然二肽化合物,具有多种生物学作用。肌肽在脑内可代谢为组氨酸并通过组氨酸脱羧酶(HDC)作用生成组胺。本实验采用永久性结扎小鼠单侧颈总动脉的血管性痴呆模型,研究血管性痴呆疾病的发病机制以及肌肽对的保护作用及其机制是否与组胺相关。方法将8周龄的C57小鼠随机分为假手术组、模型组、肌肽和组氨酸组。术前半小时腹腔注射肌肽、组氨酸,假手术组和模型组给予生理盐水,永久性结扎单侧颈总动脉建立血管性痴呆模型后饲养28天,期间隔天给予肌肽、组氨酸或生理盐水。在HDC敲除(HDC-KO)小鼠上同法制作此模型,并同法给予肌肽(200 mg/kg)。采用运动量监测、新事物识别、被动穿梭箱、水迷宫模型检测小鼠行为学指标,应用免疫染色检测小鼠大脑白质纤维密度损伤、星型胶质细胞标记物GFAP和小胶质细胞标记物Iba-1的表达,检测手术后1天和28天脑内ROS含量。结果与假手术组相比,新事物识别模型中,手术组的识别分数显著降低,各个剂量的肌肽均能显著提高术后的识别分数;被动穿梭箱模型中,手术后动物进入暗箱的潜伏期显著缩短,其中肌肽(200 mg/kg)和肌肽(500 mg/kg)组能显著延长潜伏期;水迷宫模型中,手术组学习记忆的获得与再获得能力显著降低,而肌肽(200 mg/kg)和肌肽(500 mg/kg)组均能显著改善动物的学习记忆的能力。同时,肌肽(200 mg/kg)也能显著改善HDC-KO小鼠术后在新事物识别、被动穿梭箱及水迷宫模型中的学习记忆能力,而组氨酸各剂量组在野生型小鼠上对以上指标均无明显作用。另外,此模型导致大脑白质进行性损伤,缺血后28天时大脑白质纤维密度显著疏松;缺血后14天起大脑白质区域星形胶质细胞开始激活持续到28天;同时缺血后大脑白质区域和纹状体区域小胶质细胞持续激活。肌肽能剂量依赖的抑制大脑白质纤维疏松并减少大脑白质区域星形胶质细胞的激活,并且显著抑制大脑白质区域和纹状体区域小胶质细胞的激活,组氨酸各剂量组对纤维疏松和星形胶质细胞的激活没有明显影响。在HDC-KO小鼠上,肌肽对以上指标也有显著改善作用。肌肽还可以抑制缺血后1天脑内ROS的增加。结论本实验证实了单侧颈总动脉结扎所致的皮层下缺血性血管性痴呆会导致认知功能障碍,大脑白质纤维进行性损伤、小胶质细胞的持续性激活和星形胶质细胞在后期的激活。肌肽对于皮层下缺血性血管性痴呆有明显保护作用,其机制可能不是通过转化为组胺,而可能是与抑制胶质细胞的激活和抑制ROS的产生有关。

关键词：肌肽皮层下缺血性血管性痴呆白质纤维胶质细胞

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CMT2L型的转基因小鼠模型的行为学、电生理学和病理学分析

CMT2L型的转基因小鼠模型的行为学、电生理学和病理学分析

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者：张如旭李小波资晓宏黄顺祥张付峰夏昆潘乾唐北沙中南大学湘雅三医院神经内科中南大学湘雅医院神经内科中南大学医学遗传学国家重点实验室

背景腓骨肌萎缩症(Charcot-Marie-Tooth disease,CMT)是最常见的遗传性周围神经病,具有高度的临床和遗传异质性,临床上以肢体远端肌肉进行性对称性的萎缩、无力、腱反射减弱和感觉减退等为主要特征,发病率约为1/2500。2004年,本研究小... 详细信息

背景腓骨肌萎缩症(Charcot-Marie-Tooth disease,CMT)是最常见的遗传性周围神经病,具有高度的临床和遗传异质性,临床上以肢体远端肌肉进行性对称性的萎缩、无力、腱反射减弱和感觉减退等为主要特征,发病率约为1/2500。2004年,本研究小组成功将一个来自中国湖南和湖北的常染色体显性遗传的CMT2型大家系定位于12q24并命名为CMT2L型(OMIM号608673),2005年通过位置候选克隆发现HSPB8为CMT2L型的致病基因,前期工作中我们应用微纤维注射技术构建HSPB8基因K141N的转基因小鼠模型,已获得4只HSPB8首建鼠,并顺利传代获得F2代pCAGGS-HA-HSPB8小鼠4只,F3代pCAGGS-HA-HSPB8小鼠9只。目的对已构建的pCAGGS-HA-HSPB8小鼠进行行为学、电生理学和病理学分析,以期明确HSPB8转基因小鼠模型能否良好再现CMT2L临床表型和可靠用于HSPB8生物学功能和CMT2L发病机理研究。方法取已基因鉴定的pCAGGS-HA-HSPB8转基因鼠(F2代4只,F3代9只)和正常野生型小鼠(normaltype,NT)(F2代9只,F3代9只),共4组,进行包括足迹实验、固定棒试验、旋转棒试验、痛阈实验的行为学分析;使用BL-420E+生物机能实验系统仪进行电刺激后进行坐骨神经复合肌肉动作电位振幅(CMAP)、潜伏期及运动神经传导速度(MNCV)测定的电生理学分析;坐骨神经半薄切片甲苯胺蓝染色光镜下观察和超薄切片电镜下进行周围神经病理学分析。结果足迹试验中HSPB8转基因鼠存在明显步态异常,提示后肢周围神经受损,固定棒试验中发现HSPB8转基因鼠多数蜷曲不动或行走时后肢在前肢的拖拽下费力前进,表明其运动协调能力差,HSPB8转基因鼠在旋转棒上停留时间明显少于正常小鼠,表明其神经肌肉协调能力较正常小鼠差,足底电击痛阈实验未发现HSPB8转基因鼠周围感觉神经受累。以上行为学分析HSPB8转基因鼠说明后肢周围运动神经受累,感觉神经功能相对保存;电生理学分析结果发现HSPB8转基因鼠近端和远端CMAP波幅较NT小鼠低,运动传到速度无统计学差异,表明周围神经损伤以轴索病变为主;病理学分析结果显示突变小鼠半薄切片显示薄髓纤维和无髓纤维明显增多,可见再生簇,电镜下绝大多数板层结构崩解,板层结构消失,轴索裸露和水肿变性。结论 HSPB8转基因小鼠模型良好再现了CMT2L临床表型,能可靠用于HSPB8生物学功能和CMT2L发病机理研究,这也是国际上首次成功建立的CMT2L转基因小鼠模型和HSPB8过表达转基因小鼠模型。

关键词：腓骨肌萎缩症小分子热休克蛋白 B8转基因小鼠

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